Establishment of an in vitro co-infection model of Cryptosporidium parvum and Giardia duodenalis

Abstract Background The two intestinal protozoan parasites Giardia duodenalis and Cryptosporidium parvum cause infections in a wide spectrum of vertebrates and have also been shown to infect suitable hosts simultaneously. To investigate potential effects between these parasites and on host cells, a...

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Main Authors: Manuela Kirchner, Arwid Daugschies, Cora Delling
Format: Article
Language:English
Published: BMC 2025-07-01
Series:Parasites & Vectors
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Online Access:https://doi.org/10.1186/s13071-025-06926-5
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author Manuela Kirchner
Arwid Daugschies
Cora Delling
author_facet Manuela Kirchner
Arwid Daugschies
Cora Delling
author_sort Manuela Kirchner
collection DOAJ
description Abstract Background The two intestinal protozoan parasites Giardia duodenalis and Cryptosporidium parvum cause infections in a wide spectrum of vertebrates and have also been shown to infect suitable hosts simultaneously. To investigate potential effects between these parasites and on host cells, a co-infection model with IPEC-J2 cells was established. Methods Optimal infection conditions and several infection doses of both parasites were tested. The effect of Giardia growth medium on IPEC-J2 cells was analyzed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction assay, while the effect of different infection doses of each parasite on host cell viability was investigated by CellTiter Blue cell viability assay. For co-infection, IPEC-J2 cells were first infected with C. parvum sporozoites, and 3.5 h later, G. duodenalis trophozoites were added. Parasite propagation during single infection and co-infection were analyzed by quantitative real-time polymerase chain reaction (qPCR) as well as immunofluorescent staining. Results The infection with C. parvum sporozoites had no significant impact on cell viability, while G. duodenalis trophozoites affected cell culture in a dose dependent manner. The amount of gene copies of C. parvum in single and co-infected cells did not differ significantly, while statistically higher amounts of G. duodenalis gene copies in co-infected cell cultures were identified. Conclusions In this study, single infections and co-infections of IPEC-J2 cells with C. parvum and G. duodenalis were established and optimized over a period of 72 h. Graphical Abstract
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spelling doaj-art-555c9532a2c34ab18dc1df53ac4d1fa22025-08-20T03:45:48ZengBMCParasites & Vectors1756-33052025-07-0118111410.1186/s13071-025-06926-5Establishment of an in vitro co-infection model of Cryptosporidium parvum and Giardia duodenalisManuela Kirchner0Arwid Daugschies1Cora Delling2Institute of Parasitology, Faculty of Veterinary Medicine, Leipzig UniversityInstitute of Parasitology, Faculty of Veterinary Medicine, Leipzig UniversityInstitute of Parasitology, Faculty of Veterinary Medicine, Leipzig UniversityAbstract Background The two intestinal protozoan parasites Giardia duodenalis and Cryptosporidium parvum cause infections in a wide spectrum of vertebrates and have also been shown to infect suitable hosts simultaneously. To investigate potential effects between these parasites and on host cells, a co-infection model with IPEC-J2 cells was established. Methods Optimal infection conditions and several infection doses of both parasites were tested. The effect of Giardia growth medium on IPEC-J2 cells was analyzed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction assay, while the effect of different infection doses of each parasite on host cell viability was investigated by CellTiter Blue cell viability assay. For co-infection, IPEC-J2 cells were first infected with C. parvum sporozoites, and 3.5 h later, G. duodenalis trophozoites were added. Parasite propagation during single infection and co-infection were analyzed by quantitative real-time polymerase chain reaction (qPCR) as well as immunofluorescent staining. Results The infection with C. parvum sporozoites had no significant impact on cell viability, while G. duodenalis trophozoites affected cell culture in a dose dependent manner. The amount of gene copies of C. parvum in single and co-infected cells did not differ significantly, while statistically higher amounts of G. duodenalis gene copies in co-infected cell cultures were identified. Conclusions In this study, single infections and co-infections of IPEC-J2 cells with C. parvum and G. duodenalis were established and optimized over a period of 72 h. Graphical Abstracthttps://doi.org/10.1186/s13071-025-06926-5CryptosporidiumGiardiaCell culture techniquesCo-infection
spellingShingle Manuela Kirchner
Arwid Daugschies
Cora Delling
Establishment of an in vitro co-infection model of Cryptosporidium parvum and Giardia duodenalis
Parasites & Vectors
Cryptosporidium
Giardia
Cell culture techniques
Co-infection
title Establishment of an in vitro co-infection model of Cryptosporidium parvum and Giardia duodenalis
title_full Establishment of an in vitro co-infection model of Cryptosporidium parvum and Giardia duodenalis
title_fullStr Establishment of an in vitro co-infection model of Cryptosporidium parvum and Giardia duodenalis
title_full_unstemmed Establishment of an in vitro co-infection model of Cryptosporidium parvum and Giardia duodenalis
title_short Establishment of an in vitro co-infection model of Cryptosporidium parvum and Giardia duodenalis
title_sort establishment of an in vitro co infection model of cryptosporidium parvum and giardia duodenalis
topic Cryptosporidium
Giardia
Cell culture techniques
Co-infection
url https://doi.org/10.1186/s13071-025-06926-5
work_keys_str_mv AT manuelakirchner establishmentofaninvitrocoinfectionmodelofcryptosporidiumparvumandgiardiaduodenalis
AT arwiddaugschies establishmentofaninvitrocoinfectionmodelofcryptosporidiumparvumandgiardiaduodenalis
AT coradelling establishmentofaninvitrocoinfectionmodelofcryptosporidiumparvumandgiardiaduodenalis